"Cell Disk" DNA Storage System Capable of Random Reading and Rewriting.

DNA has emerged as an appealing material for information storage due to its great storage density and durability. Random reading and rewriting are essential tasks for practical large-scale data storage. However, they are currently difficult to implement simultaneously in a single DNA-based storage system, strongly limiting their practicability. Here, a "Cell Disk" storage system is presented, achieving high-density in vivo DNA data storage that enables both random reading and rewriting. In this system, each yeast cell is used as a chamber to store information, similar to a "disk block" but with the ability to self-replicate. Specifically, each genome of yeast cell has a customized CRISPR/Cas9-based "lock-and-key" module inserted, which allows selective retrieval, erasure, or rewriting of the targeted cell "block" from a pool of cells ("disk"). Additionally, a codec algorithm with lossless compression ability is developed to improve the information density of each cell "block". As a proof of concept, target-specific reading and rewriting of the compressed data from a mimic cell "disk" comprising up to 105 "blocks" are demonstrated and achieve high specificity and reliability. The "Cell Disk" system described here concurrently supports random reading and rewriting, and it should have great scalability for practical data storage use.

[Gene edited animal models applied in human disease research].

Recently, with the development and the continuous improvement of various CRISPR systems represented by CRISPR/Cas9, gene editing technology has been gradually improved, and widely applied to the preparation of animal models of human diseases. The gene edited animal models provide important materials for the study of pathogenesis, pathological process, prevention and treatment of human diseases. At present, the gene edited animal models used in human disease research include mainly the rodent models represented by mice and rats, and large animal models represented by pigs. Among them, rodents differ greatly from humans in all aspects of their bodies and have short life span as well, which cannot provide effective evaluation and long-term tracking for the research and treatment of human diseases. On the other hand, pig is closer to human in physiology, anatomy, nutrition and genetics, which provides an important animal model in the field of organ transplantation and human disease research. In this paper, the application of the gene edited animal models was summarized in the researches of 5 human diseases such as neurodegenerative diseases, familial hypertrophic cardiomyopathy, cancer, immunodeficiency diseases and metabolic diseases. We hope this paper will provide a reference for the research of human diseases and the preparation of relative animal models.

Prostate stem cell antigen mRNA expression in preoperatively negative biopsy specimens predicts subsequent cancer after transurethral resection of the prostate for benign prostatic hyperplasia.

BACKGROUND: Recent data showed that prostate stem cell antigen (PSCA) mRNA expression in transurethral resection of the prostate (TURP)-resected tissues predicted the subsequent prostate cancer after TURP in benign prostatic hyperplasia (BPH) patients with both PSA < 4.0 ng/ml and normal DRE findings. This study was to determine whether PSCA mRNA positivity in preoperatively negative prostatic biopsy samples from BPH men with PSA > 4.0 ng/ml and/or suspicious DRE findings had predictive performance following TURP. MATERIALS AND METHODS: PSCA in situ hybridization was performed on negative prostatic biopsies taken before TURP from 166 enrolled symptomatic BPH patients, who were continuously followed for 5 years postoperatively. Predictive performance of PSCA mRNA for subsequent cancer onset was evaluated by univariate and multivariate Cox proportional hazards models with bootstrapping and concordance indices. RESULTS: PSCA mRNA was detected in 42/166 (25.3%) of the preoperatively negative biopsy specimens, with a mean positive-labeling cells of 31.6%, in which 31 patients were identified as having subsequent PCa on follow-up. Of 124 patients with negative expression for PSCA mRNA none were subsequently diagnosed with PCa. The examination of Spearman's rank correlation coefficient showed that PSCA mRNA expression levels were positively and statistically correlated with higher Gleason score (r = 0.88, P < 0.001) and clinical T stage (r = 0.84, P < 0.001). A final multivariate Cox proportional hazards model demonstrated that only PSCA mRNA expression in negative prostatic biopsies was predictive of the subsequent cancer development after TURP (hazard ratio = 3.49; 95% CI: 2.02-4.75; P < 0.001), with the concordance index of 0.893. CONCLUSIONS: This prospective study identifies PSCA mRNA in preoperatively negative prostatic biopsies as a significant predictor of subsequent cancer after TURP.

Heat shock protein 70 antisense oligonucleotide inhibits cell growth and induces apoptosis in human gastric cancer cell line SGC-7901.

AIM: Heat shock protein (HSP)70 is over-expressed in human gastric cancer and plays an important role in the progression of this cancer. We investigated the effects of antisense HSP70 oligomer on human gastric cancer cell line SGC-7901, and its potential role in gene therapy for this cancer. METHODS: Human gastric cancer cell line SGC-7901 was treated in vitro with various concentrations of antisense HSP70 oligonucleotides at different intervals. Growth inhibition was determined as percentage by trypan blue dye exclusion test. Extracted DNA was electrophoresed on agarose gel, and distribution of cell cycle and kinetics of apoptosis induction were analyzed by propidium iodide DNA incorporation using flow cytometry, which was also used to detect the effects of antisense oligomer pretreatment on the subsequent apoptosis induced by heat shock in SGC-7901 cells. Proteins were extracted for simultaneous measurement of HSP70 expression level by SDS-PAGE Western blotting. RESULTS: The number of viable cells decreased in a dose- and time-dependent manner, and ladder-like patterns of DNA fragments were observed in SGC-7901 cells treated with antisense HSP70 oligomers at a concentration of 10 mumol/L for 48 h or 8 micromol/L for 72 h, which were consistent with inter-nucleosomal DNA fragmentation. Flow cytometric analysis showed a dose- and time-dependent increase in apoptotic rate by HSP70 antisense oligomers. This response was accompanied with a decrease in the percentage of cells in the G1 and S phases of the cell cycle, suggesting inhibition of cell proliferation. In addition, flow cytometry also showed that pretreatment of SGC-7901 cells with HSP70 antisense oligomers enhanced the subsequent apoptosis induced by heat shock treatment. Western blotting demonstrated that HSP70 antisense oligomers inhibited HSP70 expression, which preceded apoptosis, and HSP70 was undetectable at the concentration of 10 micromol/L for 48 h or 8 micromol/L for 72 h. CONCLUSION: Antisense HSP70 oligomers can abrogate HSP70 expression in SGC-7901 cells, which may in turn induce apoptosis and inhibit cell proliferation, conversely suggesting that HSP70 is required for the proliferation and survival of human gastric cancer cells under normal conditions.

Inhibition of transfected PTEN on human colon cancer.

AIM: To study the inhibitory effect of transfected PTEN on LoVo cells. METHODS: Human PTEN cDNA was transferred into LoVo cells via lipofectin and PTEN mRNA levels and its expression were analyzed by Western blot and flow cytometry. Before or after transfection, the effects of 5-Fu on inhibiting cell proliferation and inducing apoptosis were measured by flow cytometry, DNA bands and MTT. RESULTS: PTEN transfection significantly up-regulated PTEN expression in LoVo cells. 5-Fu inhibited cell proliferation and induced apoptosis in transfected LoVo cells. CONCLUSION: Transfected PTEN can remarkably up-regulate PTEN expression in LoVo cells and promote the apoptosis. PTEN transfection is associated with 5-Fu treatment effect and has a cooperatively cytotoxic effect.

Effect of pharmacologic preconditioning with tetrandrine on subsequent ischemia/reperfusion injury in rat liver.

The effect of tetrandrine (TET) pretreatment of Wistar rats subjected to warm hepatic ischemia/reperfusion (I/R) was investigated. After 50 minutes of ischemia in the left and median lobes of the liver and 24 hours of reperfusion (I/R group), the rats were killed. The TET+I/R group rats were pretreated with TET (50 mg/kg body weight IP) 30 minutes prior to the onset of ischemia. Blood samples were taken for measurement of alanine aminotransferase (ALT), aspartate aminotransferase (AST), and lactate dehydrogenase (LDH). Tissue was taken from the ischemic lobes for measurement of superoxide dismutase (SOD), malonyldialdehyde (MDA), and myeloperoxidase (MPO); determination of the wet/dry weight (W/D) ratio; and histologic studies. The results showed that ALT, AST, and LDH levels in serum were increased in the I/R group; tissue MDA generation, MPO activity, and the W/D ratio were also increased, accompanied by decreased SOD activity. The serum ALT, AST, and LDH levels, as well as the tissue MPO level and W/D ratio, were lower in the TET+ I/R group than in the I/R group; and the SOD level was higher in the TET+IR group than in the I/R group. Moreover, the serum ALT and AST, tissue MDA, and W/D ratio in the TET+I/R group were higher, and the SOD was lower than in the sham group. The histologic examination showed protection against liver damage in the TET+I/R group. The results demonstrated that pretreatment with TET could somewhat protect the liver against I/R injury but does not prevent it. The simultaneous decrease of both lipid peroxide generation and polymorphonuclear neutrophil infiltration in the ischemic liver may explain the acquisition of tolerance following administration of TET.